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[在与核仁DNA结合的情况下,用四氟铝酸盐处理同步化的K562细胞不会调节溴化乙锭和4',6-二脒基-2-苯基吲哚的荧光]

[Treatment of synchronized K562 cells by tetrafluoroaluminate does not modulate the fluorescence of ethidium bromide and 4'6-diamidine-2-phenylindole in case of the binding with nucleoid DNA].

作者信息

Anisimov A G, Bolotnikov I A

机构信息

Department of Biochemistry, Petrozavodsk State University.

出版信息

Tsitologiia. 1999;41(8):680-4.

Abstract

Earlier we showed that 4-hours treatment of cells K562 with the GTP-binding protein activator AlF4- (10 mM NaF + 20 microM AlCl3) increased the DNA fragmentation on an average to 5% of the total 3H-thymidine-labeled DNA. The viability of cells under these conditions did not change. It has been suggested that gene toxic action of AlF4- is a result of cell proliferation block induced by AlF4-. In the present work we tried to determine possible changes in the ethidium bromide and 4',6-diamidine-2-phenylindole (DAPI) fluorescence when they bind with nucleoid DNA of synchronized cells K562 treated with AlF4-. Cells K562 were incubated for synchronization with 2 mM thymidine during 15 hours. Under these conditions DNA synthesis (3H-thymidine uptake) was suppressed by 94-99%. It has been found that the treatment of "cool" thymidine-incubated cells K562 with AlF4- did not change the fluorescence of either ethidium or DAPI. The presence of phorbol-12-myristate-13-acetate (PMA) in the incubation medium did not influence the results. On the other, hand the rat thymocytes incubated with dexametazone (2 microM) during 4 hours (positive control of DNA fragmentation) demonstrated the increase in both parameters. PMA decreased the ethidium fluorescence that correspond to its (PMA) ability to suppress fragmentation of thymocyte DNA. On the base of the results we suggested that AlF4- did not induce DNA fragmentation in the cells K562 with the blocked DNA synthesis.

摘要

我们之前的研究表明,用GTP结合蛋白激活剂AlF4-(10 mM NaF + 20 microM AlCl3)处理细胞K562 4小时,可使DNA片段化平均增加至总3H-胸腺嘧啶标记DNA的5%。在这些条件下细胞的活力没有变化。有人提出,AlF4-的基因毒性作用是由AlF4-诱导的细胞增殖阻滞所致。在本研究中,我们试图确定溴化乙锭和4',6-二脒基-2-苯基吲哚(DAPI)与经AlF4-处理的同步化细胞K562的类核DNA结合时可能发生的变化。将细胞K562与2 mM胸腺嘧啶一起孵育15小时以进行同步化。在这些条件下,DNA合成(3H-胸腺嘧啶摄取)被抑制了94%-99%。已发现用AlF4-处理经“冷”胸腺嘧啶孵育的细胞K562,不会改变溴化乙锭或DAPI的荧光。孵育培养基中佛波醇-12-肉豆蔻酸酯-13-乙酸酯(PMA)的存在不影响结果。另一方面,用地塞米松(2 microM)孵育4小时的大鼠胸腺细胞(DNA片段化的阳性对照)显示这两个参数均增加。PMA降低了溴化乙锭的荧光,这与其(PMA)抑制胸腺细胞DNA片段化的能力相符。基于这些结果,我们认为AlF4-不会在DNA合成受阻的细胞K562中诱导DNA片段化。

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